AIM: To investigate the effectiveness of recombinanted retrovirus vector in gene therapy. METHODS: The retroviral vector pLXSN S was constructed and transferred into PA317 by means of electroporation, then HepG 2?RAW264.7 and EL 4 cells were infected with the pseudovirus produced from PA317, which highly expressed HBsAg . HBsAg expression was tested by RT PCR and ELISA. RESULTS: HBsAg was expressed variously in the eukaryotic cells mentioned above. HBsAg ( A value) of the cell supernatants (48 hours) were 0.92,0.53,0.42, respectively. CONCLUSION: The vector used in this study was an effective vector to carry genes of interest to target cells and macrophage, and high level HBsAg was expressed in antigen presenting cell such as macrophage, It indicated that plasmid immunity can induce the B lymphocyte and T lymphocyte response to hepatitis B virus surface antigen by stimulating macrophage. As a vector, it may be useful in the test for gene immunity and gene therapy.

Objective To study the effects of matrine on accumulation of ? globin mRNA and induction of erythroid differentiation in K562 cells in vitro.Methods K562 cells were cultured for 6 days with different concentration of matrine,viable cell counts were determined by trypan-blue dye exdusion test. Erythroid differentiation was evaluated by percentage of benzidine-positive cells at different days after culture. Morphological changes were observed under microscope after Wright-Gimesa staining; ? globin mRNA was quantitative by real time quantitative reverse transcript polymerase chain reaction(RT-PCR).Results Different concentrations of matrine inhibited proliferation of K562 cells in dose-dependent manner; otherwise, K562 cells were successfully induced by erythroid differentiation with matrine. After treatment with matrine, percentage of benzidine-positive cells significantly increased from 0.7% to 15.7% and characteristic changes of erythroid differentiation in the cell morphology were observed, G? globin mRNA had a preferential increase (2.7 fold)in K562 cells. Conclusions Matrine accumulation G? globin mRNA and induced erythroid differentiation of K562 cells. The results provides an experimental evidence for the pharmacological therapy of hematological diseases associated with a failure in the expression of normal ? globin genes.

Objective To evaluate the pharmacokinetics of remifentanil in adult patients undergoing general anesthesia.Methods Ten ASA Ⅰ or Ⅱ adult patients of either sex undergoing elective surgery under general anesthesia were enrolled in this study.The patients were premedicated with atropine 0.5 mg.Anesthesia was induced with fentany1-2 ?g?kg~(-1),lidocaine 0.5-1.0 mg?kg~(-1),propofol 1.5-2.0 mg?kg~(-1) and vecuronium 0.1 mg?kg~(-1) and maintained with isoflurane -O_2 inhalation and intermittent i.v.boluses of vecuronium after tracheal intubation.The patients were mechanically ventilated.P_(ET)CO_2 was maintained at 30-40 mm Hg. Remifentanil 5 ?g?kg~(-1) was injected i.v.over lmin after induction of anesthesia.Arterial blood samples were taken before(baseline)and at 1,2,3,5,7,10,15,20,25,30,45,60 and 90 rain after drug administration for determination of blood remifentanil concentration using high performance liquid chromatography/mass spectrometry. The data were analyzed using 3P87 pharmacokinetic program.Results The pharmacokinetic profile of remifentanil was best described by a “two-compartment” model.The distribution half-life(t_(1/2?))(1.6?0.5)min,the elimination half-life(t_(1/2?))(22?10)min,total clearance(CL)(2.1?0.4)L/min and the volume of distribution (V_d)(66?29)L.Conclusion The volume of distribution calculated in this study is significantly different from that reported in previous studies,indicating that there may be some difference in the pharmacokineties of remifentanil between different populations.

In order to study the accumulation of Fritillaria thunbergii cultivar, peimine content in Xiaye, Kuanye, Duozi and Xiaosanzi bulbs of different sizes and parts was determined by HPLC-ELSE. The results indicated that the peimine content varied significantly with the cultivar type, the size and part of bulb. The distribution laws of peimine were as follow: Xiaosanzi > Duozi > Xiaye > Kuanye, small-size bulb > big-size bulb, core bud > scale. The peimine yield per plant in Duozi was the highest.
Cevanes ; analysis ; Chromatography, High Pressure Liquid ; Fritillaria ; chemistry ; growth & development

Objective To lessen the learning curve of percutaneous renal lithotripsy(PCNL),we introduced a novel ex-vivo learning and training model for PCNL under fluoroscopy and ultrasonography-guided access. Methods The model was set up nailing an adult porcine kidney with＞3 cm ureter (freshly removed from the slaughtering factory),with a full thickness flap of the thoracic wall or abdomen wall with two ribs,to a board.The porcine kidney was placed within the flap with the catheterized ureter outside.The kidney was enclosed by the flap so as to create a roodel for percutaneous renal surgery;with the ribs overlying the upper portion of the kidney.The model was fixed to the board by two nails.Artificial stone material was implanted in the renal pelvis.Fluoroscopy guidance access:Retrograde pyelography via injection of contrast medium into the ureteral catheter images the collecting system. After the long axis of the target calyx is identified,the C-arm is rotated 30 degrees toward the surgeon,placing the C-arm axis in the same posterior plane of the kidney.The needle is advanced in the plane of the fluoroscopic beam,and the appropriate needle placement is determined by obtaining a bull's-eye sign on the fluoroscope screen. Rotating the C-arm to a vertical position monitors the depth of the needle penetration. Ultrasonography guidance access:The renal pelvis can he filled with normal saline through a catheter to simulate hydronephrosis and the target calyx is identified under ultrasonography guidance.The tract dilation and stone disintegration were followed.After training,the kidney can be opened to examine the target calyx and the complication of dilation. Results Altogether,126 urologists attended a urologic endoscopic technique training course.Of the 126 trainees,104 (82.5%) successfully performed the whole percutaneous procedure.At the end of training,114 (90.5 %) of the 126 attendees rated the porcine kidney model for simulation of percutaneous renal surgery as very helpful or helpful.Conclusions This biological training model simulates realistically the clinical procedure of PCNL with respect to trainee experience in a low stress environment that provides an opportunity for repetitive performances in order to learn basic technical skills for the clinical procedure of PCNL in the future.

Objective To investigate the value of temporary balloon occlusion of the abdominal aorta in the treatment of complete placenta previa with placenta accreta. Methods From January 2015 to February 2016, 24 cases of complete placenta previa with placenta accreta were treated with temporary balloon occlusion of the abdominal aorta (the study group) before cesarean, and 24 cases of complete placenta previa with placenta accreta did not receive balloon occlusion (the control group). The operation time, intraoperative blood loss, intraoperative blood transfusion volume, the perioperative hemoglobin level, the hysterectomy rate and the related complications were compared retrospectively.Also, the hospitalization time, the blood coagulation parameters after operation, including activated partial thromboplastin time (APTT), fibrinogen (FIB), D-Dimer and reperfusion injury parameters including creatine phosphokinase (CK), creatine phosphokinase isoenzyme (CK-MB), lactate dehydrogenase (LDH) and serum creatinine were compared between the 2 groups. Results The blood loss [750 ml (400-2 000 ml) vs 2 000 ml (1 500-2 375 ml);Z=-3.214, P=0.001] and blood transfusion volume [200 ml (0-800 ml) vs 800 ml (0-1 200 ml);173, P=0.030] in the study group were lower than in the control group. The hemoglobin difference between before and after operation in the study group was lower than the control group [(12.8±13.4) g/L vs (22.9±20.1) g/L;t=-2.041, P=0.047]. In the study group, there were still bleeding in 13 cases after releasing the balloon, 5 of them received uterine artery embolization, 5 cases received uterine artery ligation, and 3 cases received uterine packing. One case had venous thrombosis in the right lower limb. Two cases (8%,2/24) in the control group had hysterectomy, while none in the study group, there was no statistical significance (P=0.489). Conclusions Temporary balloon occlusion of the abdominal aorta can effectively reduce blood loss and blood transfusion in the treatment of complete placenta previa with placenta accreta, but there is still the risk of continuing bleeding after releasing the balloon. Other methods of hemostasis might be needed.

Objective:To study the regulatory effects of Ligustrazine Hydrochloride(LHC)on tumor-induced immunosuppression by Colon26 cells in vitro.Methods:Colon26 cells were cultured for 48 h in the presense of LHC and either the cell fraction or the cultural supernatants was collected,with the untreated Colon26 cells as control for the study.The down-regulating effects of LHC on tumor immunosuppressions (including the suppressed NK killing and ConA induced transformation of murine spleen cells detected by MTT,and the reduced expression levels of IL-2R?,CD3?+?+ and CD3?-?+ detected by FCM) were determined.The concentrations of immunosuppressive cytokines,including TGF-?1,VEGF,IL-4,IL-6 and IL-10,in the supernatants were analyzed by quantitative ELISA.The relationship among the down-regulatory effects of LHC on secretion immunosuppressive cytokines and tumor immunosuppressions were evaluated by multiple linear regression analysis.Results:All of the cytokines assayed were found in the supernatant of Colon26 treated without LHC,in which TGF-?1 was the highest,and the significant inhibition of five immune functions mentioned above was showed.To the Colon26 treated by LHC,the concentrations of TGF-?1,IL-6 and IL-10 in the first re-cultured supernatant and its inhibition of five immunol functions decreased greatly.The concentrations of TGF-?1 and IL-6 in the second re-cultured supernatant and its inhibitions of transformation,CD3+?+ and CD3-?+resumed highly.The positive correlations existed between TGF-?1 and inhibition of immunol functions except for transformation,between IL-6 and inhibition of transformation or CD3-?+,between IL-10 and inhibition of NK killing or IL-2R? or CD3+?+,respectively.Conclusion:LHC can exert down-regulate effects on Colon26 secretion of immunosuppressors and its tumor immunosuppression.Reducing tumor immunosuppression of Colon26 through decreasing its secretion of immunosuppressors should be one of anti-tumor mechanisms of LHC.

Objective To observe the effects of five flavonoids include rutin(RU),dihydromyricetin(DMY),hesperetin(HP),daidzein(DA)and hydroxysaffor yellow A(HYSA)on myocardiocyte apoptosis induced by H2O2 and to explore their relationships with Keshan disease and the possible mechanism.Methods Primary cultured cardiocytes of neonatal rats were randomly divided into control group,model group,and flavonoids preincubation group.The cardiocyte apoptosis was examined by fluorescent staining,the rates of apoptosis were detected by flow cytometry,the expression of Bcl-2 family proteins associated with apoptosis were observed:by Western blot.Results Compared with model group[(24.33±6.51)%],RU[(13.95±3.80)%],DA[(11.82±3.50)%],HYSA[(12.33±3.78)%]could decreased the rate of apoptosis(P＜0.05).The five flavonoids could up-regulate Bcl-2 expression,down-regulate Bax expression,and increase the Bcl-2/Bax ratio[RU(0.989±0.094),DMY(0.931±0.280),HP(0.980±0.095),DA(1.049±0.092),HYSA(1.031±0.039),vs model(0.490±0.046),the difference had statistical significances(P＜0.05)],but the Bcl-xl did not significantly changed(P＞0.05).Conclusions RU,DMY,HP,DA and HYSA have antiapoptotic effects on cardiomyocyte via regulating Bcl-2 and Bax,which gives us a hint in prevention and treatment of Keshan disease.

0.05).Conclusions SNP of 2350G→A in ACE gene is associated with MI,AA genotype is probably a genetic marker of MI in Han nationality.